The cholesterol concentration of uncooked and cooked longissimus muscle, uncooked subcutaneous fat and serum, as affected by days on feed, breed type and sex class in a factorial arrangement, and the relationship between serum cholesterol and tissue cholesterol were studied. Days on feed, breed type and sex class had no effect (P greater than .05) on tissue cholesterol concentrations. Overall least-squares means for uncooked and cooked longissimus muscle and subcutaneous fat were 63.32, 80.27 and 98.90 mg of cholesterol/100 g of tissue, respectively. Cholesterol concentration was 26.8% higher in cooked vs uncooked muscle and 56.2% higher in uncooked subcutaneous fat compared with uncooked muscle. Serum cholesterol concentration was lower (P less than .05) for fullblood Chianinas and steers (122.6 and 133.9 mg/dl) compared with crossbreds and heifers (162.3 and 151.0 mg/dl), respectively. Serum cholesterol concentration showed a cubic response (P less than .05) over days on feed; it was lowest at 0 d (79.6 mg/dl), increased up to 77 d (156.3 mg/dl), leveled off (154.7 mg/dl), then increased again (179.3 mg/dl). Serum cholesterol had low and insignificant correlation coefficients with all tissue cholesterol concentrations (r = .08 to .22; P greater than .05). Results indicate that muscle and fat tissue cholesterol concentrations do not vary in response to the differences in breed type, sex class or time on feed represented in the present study and thus may be an inherent characteristic dependent upon quantitative needs for cellular membrane functions. However, serum cholesterol concentration was affected by treatments evaluated in this study and may be related to factors affecting lipid metabolism.